Limits...
A Novel Mutation of DAX-1 Associated with Secretory Azoospermia.

Mou L, Xie N, Yang L, Liu Y, Diao R, Cai Z, Li H, Gui Y - PLoS ONE (2015)

Bottom Line: A number of coding mutations unique to the patient group, including two synonymous mutations and six missense mutations, were identified.Of the missense mutations, our functional assay demonstrated that the V385L mutation caused the reduced functioning of DAX-1.This novel mutation (p.

View Article: PubMed Central - PubMed

Affiliation: Shenzhen Key Laboratory of Genitourinary Tumor, Shenzhen Domesticated Organ Medical Engineering Research and Development Center, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518035, China; Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics, Institute of Urology, Peking University Shenzhen Hospital, Biomedical Research Institute, Shenzhen PKU-HKUST Medical Center, Shenzhen, 518036, China.

ABSTRACT
Secretory azoospermia is a severe form of male infertility caused by unknown factors. DAX-1 is predominantly expressed in mammalian reproductive tissues and plays an important role in spermatogenesis because Dax-1 knockout male mice show spermatogenesis defects. To examine whether DAX-1 is involved in the pathogenesis of secretory azoospermia in humans, we sequenced all of the exons of DAX-1 in 776 patients diagnosed with secretory azoospermia and 709 proven fertile men. A number of coding mutations unique to the patient group, including two synonymous mutations and six missense mutations, were identified. Of the missense mutations, our functional assay demonstrated that the V385L mutation caused the reduced functioning of DAX-1. This novel mutation (p. V385L) of DAX-1 is the first to be identified in association with secretory azoospermia, thereby highlighting the important role of DAX-1 in spermatogenesis.

No MeSH data available.


Related in: MedlinePlus

Interactions between DAX-1 mutants and AR.HeLa cells were transfected with expression vectors for AR, DAX-1 WT, DAX-1 R51K, DAX-1 C104W, DAX-1 A242V, DAX-1 E256Q, DAX-1 V385L, and DAX-1 I427V as indicated. DAX-1 and AR were immunoprecipitated (IP) with an anti-HA antibody and AR antibody, respectively. Then, the immunocomplexes were analyzed by SDS-PAGE and Western blotting (WB) analysis using anti-HA and anti-AR antibodies as indicated.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4514677&req=5

pone.0133997.g002: Interactions between DAX-1 mutants and AR.HeLa cells were transfected with expression vectors for AR, DAX-1 WT, DAX-1 R51K, DAX-1 C104W, DAX-1 A242V, DAX-1 E256Q, DAX-1 V385L, and DAX-1 I427V as indicated. DAX-1 and AR were immunoprecipitated (IP) with an anti-HA antibody and AR antibody, respectively. Then, the immunocomplexes were analyzed by SDS-PAGE and Western blotting (WB) analysis using anti-HA and anti-AR antibodies as indicated.

Mentions: DAX-1 inhibits the transcriptional activity of AR through protein-protein interactions [18–20]. To determine whether the identified missense mutations in DAX-1 affect its ability to bind to AR, WT and mutated DAX-1 constructs were transfected into HeLa cells with an AR plasmid. The results showed that all DAX-1 R51K, C104W, A242V, E256Q, and I427V mutants co-immunoprecipitated with AR similar to DAX-1 WT but that V385L was weakly bound to AR (Fig 2). Collectively, these results indicated that the V385L mutation affected the interaction between DAX-1 and AR.


A Novel Mutation of DAX-1 Associated with Secretory Azoospermia.

Mou L, Xie N, Yang L, Liu Y, Diao R, Cai Z, Li H, Gui Y - PLoS ONE (2015)

Interactions between DAX-1 mutants and AR.HeLa cells were transfected with expression vectors for AR, DAX-1 WT, DAX-1 R51K, DAX-1 C104W, DAX-1 A242V, DAX-1 E256Q, DAX-1 V385L, and DAX-1 I427V as indicated. DAX-1 and AR were immunoprecipitated (IP) with an anti-HA antibody and AR antibody, respectively. Then, the immunocomplexes were analyzed by SDS-PAGE and Western blotting (WB) analysis using anti-HA and anti-AR antibodies as indicated.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4514677&req=5

pone.0133997.g002: Interactions between DAX-1 mutants and AR.HeLa cells were transfected with expression vectors for AR, DAX-1 WT, DAX-1 R51K, DAX-1 C104W, DAX-1 A242V, DAX-1 E256Q, DAX-1 V385L, and DAX-1 I427V as indicated. DAX-1 and AR were immunoprecipitated (IP) with an anti-HA antibody and AR antibody, respectively. Then, the immunocomplexes were analyzed by SDS-PAGE and Western blotting (WB) analysis using anti-HA and anti-AR antibodies as indicated.
Mentions: DAX-1 inhibits the transcriptional activity of AR through protein-protein interactions [18–20]. To determine whether the identified missense mutations in DAX-1 affect its ability to bind to AR, WT and mutated DAX-1 constructs were transfected into HeLa cells with an AR plasmid. The results showed that all DAX-1 R51K, C104W, A242V, E256Q, and I427V mutants co-immunoprecipitated with AR similar to DAX-1 WT but that V385L was weakly bound to AR (Fig 2). Collectively, these results indicated that the V385L mutation affected the interaction between DAX-1 and AR.

Bottom Line: A number of coding mutations unique to the patient group, including two synonymous mutations and six missense mutations, were identified.Of the missense mutations, our functional assay demonstrated that the V385L mutation caused the reduced functioning of DAX-1.This novel mutation (p.

View Article: PubMed Central - PubMed

Affiliation: Shenzhen Key Laboratory of Genitourinary Tumor, Shenzhen Domesticated Organ Medical Engineering Research and Development Center, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518035, China; Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics, Institute of Urology, Peking University Shenzhen Hospital, Biomedical Research Institute, Shenzhen PKU-HKUST Medical Center, Shenzhen, 518036, China.

ABSTRACT
Secretory azoospermia is a severe form of male infertility caused by unknown factors. DAX-1 is predominantly expressed in mammalian reproductive tissues and plays an important role in spermatogenesis because Dax-1 knockout male mice show spermatogenesis defects. To examine whether DAX-1 is involved in the pathogenesis of secretory azoospermia in humans, we sequenced all of the exons of DAX-1 in 776 patients diagnosed with secretory azoospermia and 709 proven fertile men. A number of coding mutations unique to the patient group, including two synonymous mutations and six missense mutations, were identified. Of the missense mutations, our functional assay demonstrated that the V385L mutation caused the reduced functioning of DAX-1. This novel mutation (p. V385L) of DAX-1 is the first to be identified in association with secretory azoospermia, thereby highlighting the important role of DAX-1 in spermatogenesis.

No MeSH data available.


Related in: MedlinePlus